Ig DH Gene Segment Transcription and Rearrangement Before Surface Expression of the Pan-B–Cell Marker CD19 in Normal Human Bone Marrow by F.E. Bertrand, L.G. Billips, P.D. Burrows, G.L. Gartland, H. Kubagawa, and H.W. Schroeder Blood Volume 90(2):736-744 July 15, 1997 ©1997 by American Society of Hematology

Bone marrow B-lineage subpopulations collected on the basis of CD34, CD19, and IgM surface expression. Bone marrow B-lineage subpopulations collected on the basis of CD34, CD19, and IgM surface expression. (A) Cartoon illustrating each of the five subpopulations collected and the B-lineage developmental stage that each represents. Thick lines indicate higher levels of surface expression of the respective B-cell marker. (B) Typical collection gates used to separate the FBM subpopulations are shown in these FACS profiles of 22-week-old FBM stained with anti-CD34, anti-CD19, and anti-IgM as described in the Materials and Methods. (C) Typical collection gates used to separate the ABM subpopulations are shown in these FACS profiles of 30-year-old ABM stained with anti-CD34, anti-CD19, and anti-IgM as described in the Materials and Methods. (D) Typical postsort FACS analysis of the fetal CD34+ CD19− subpopulation illustrating 99% purity in each subpopulation. Postsort analysis of the other fetal and adult subpopulations collected also indicated 99% purity (data not shown). F.E. Bertrand III et al. Blood 1997;90:736-744 ©1997 by American Society of Hematology

Map of the human VH6-DJ-Cμ locus. Map of the human VH6-DJ-Cμ locus. Shown is a 400-kb region extending from VH6 through Cμ. The approximate positions of the DXP gene segments are denoted within each of the four DH tandem repeats in the distal DH locus, labeled as DXP1 (D21/7) and DXP1′ (D21/0.5), DXP2, DXP3, and DXP4. The expanded portion indicates the location of DHQ52 within the proximal DH locus, the six JH gene segments, and the intronic sequence between JH and Cμ that includes the proximal IgH enhancer. The composition of germline transcripts from DHQ52, Iμ, and germline Cμ is illustrated. For the transcripts shown, boxes indicate transcribed regions and lines indicated areas that are removed due to RNA processing. F.E. Bertrand III et al. Blood 1997;90:736-744 ©1997 by American Society of Hematology

IgH transcripts in human bone marrow B-lineage subpopulations. IgH transcripts in human bone marrow B-lineage subpopulations. Iμ, germline Cμ, germline DHQ52, and rearranged DHQ52 DJ, DXP DJ, VH3, and VH6 transcripts were detected as described in the Materials and Methods and the Results. (A) FBM IgH transcripts. (B) ABM IgH transcripts. Shown are results representative of repeated PCR analysis of four independently sorted FBM samples and three independently sorted ABM samples. F.E. Bertrand III et al. Blood 1997;90:736-744 ©1997 by American Society of Hematology

PCR assay to detect IgH DJ rearrangements in sorted FBM B-lineage subpopulations. PCR assay to detect IgH DJ rearrangements in sorted FBM B-lineage subpopulations. (A)FBM DHQ52 DJ rearrangements. (B) FBM DXP DJ rearrangements. No germline band is observed in the DXP rearrangements lane because the primers anneal to sequences that are widely separated (20 kb) in unrearranged DNA. Shown are results representative of repeated PCR analysis of three independently sorted FBM samples and two independently sorted ABM samples. F.E. Bertrand III et al. Blood 1997;90:736-744 ©1997 by American Society of Hematology

Quantitation of RT-PCR IgH products in FBM and ABM Quantitation of RT-PCR IgH products in FBM and ABM. Shown are graphs depicting the average relative abundance of individual IgH transcripts detected in the fetal (N = 3) and adult (N = 2) subpopulations. Quantitation of RT-PCR IgH products in FBM and ABM. Shown are graphs depicting the average relative abundance of individual IgH transcripts detected in the fetal (N = 3) and adult (N = 2) subpopulations. (▴) Fetal samples. (•) Adult samples. F.E. Bertrand III et al. Blood 1997;90:736-744 ©1997 by American Society of Hematology